KR101465972B1 - Composition for renewing skin containing vacuum-distillated chamomile extract as effective component - Google Patents

Abstract

The present invention relates to a composition for regenerating skin containing chamomile reduced-pressure distillation extract as an active ingredient, wherein the skin regeneration composition of the present invention containing camphor or a chamomile decompression distillate extract containing the same, And has no side effects because it contains only components derived from natural herb plants. Therefore, it can be used in various forms such as medicines, quasi-drugs, cosmetics and food materials.

Description

TECHNICAL FIELD [0001] The present invention relates to a skin regeneration composition containing a chamomile decompression distillation extract as an active ingredient,

The present invention relates to a composition for regenerating skin comprising chamomile decompression extract as an active ingredient, and more particularly to a composition for skin regeneration comprising camphor (1R) -1,7,7-trimethylbicyclo [2.2.1] heptan-2-one ) Or a composition for regenerating skin containing camomile hydrosol as an active ingredient, a cosmetic composition and a food composition for regenerating skin containing as an active ingredient camomile or a camomile hydrosol including the same, And a cosmetically acceptable salt or a camomile hydrosol comprising the above-mentioned camphor as an active ingredient.

Chamomile (Matricaria recutita L.) belongs to Asteraceae and is widely used as a herbal tea. For many years, chamomile has been used to relieve intestinal gas, headache, insomnia, back pain, neuralgia, rheumatism, skin diseases, indigestion and gout diseases. Among the various chamomile, German Chamomile is widely used and received more scientific evaluation. German chamomile contains bisabolol, which has low toxicity and sensitivity in skin care, whereas Roman Chamomile, Anobillis and Moroccan Chamomile contain bis-gallol not. Recent studies have shown that chamomile essential oils are highly effective in antimicrobial and antioxidant (Natarajan et al . , Phytotherapy Research , 27 (1) 118-125, 2013). Chamomile essential oils contain a variety of phytochemicals including flavonoids, some of which exhibit anti-inflammatory and antiallergic activity. Chamomile essential oils have long been studied in many fields, including pharmaceuticals, perfumes, cosmetics and the food industry. However, the activity of the chamomile hydrosol has not been reported. Therefore, this study aims to reveal that chamomile hydrosol shows high efficacy for skin treatment like essential oil.

Fibroblast is a fusiform cell, derived from multipotential mesenchymal cells that are involved in fiber formation involved in skin structure and elasticity. Fibroblasts play an important role in determining the skin characteristics by controlling the moisture and the extracellular matrix such as collagen, elastin, and glycoprotein. Fibroblasts also secrete matrix metalloproteinases such as collagenase and Tissue Inhibitors of Metalloproteinases. Along with the synthesis of extracellular matrix, the balance of these proteases and inhibitors regulates skin properties and extracellular matrix remodeling.

An important indicator of cell proliferation and migration is the increase in reactive oxygen species (ROS). Generation of reactive oxygen species is not only seen in damage, but also in cell growth by various stimuli. In normal physiological signals, the role of essential oxygen-dependent reactive oxygen species is to promote transcription factors such as NF-κB, MAP kinase and PI3K / AKT. Active oxygen species promote cell migration in cells and affect the action of mobile cells in non-migrating cells. Recent studies have shown that reactive oxygen species induce epithelial-mesenchymal transition important for cell migration and wound healing (DY Rhyu et al . , J Am Soc Nephrol , 16: 667, 2005). This process occurs when cells lose cell-to-cell contact and the conversion from epithelial cells to mesenchymal cells occurs, and vimentin, Slug, Snail and E-cadherin ) Is well known as a key indicator of epithelial-mesenchymal transition. The key signaling pathway that regulates cell proliferation in many cells is the PI3K and MEK / ERK pathway. The most distinctive effect of PI3K is an increase in phosphorylated AKT. Activated AKT inhibits TSC1 / TSC2 and activates mTOR by phosphorylating Raptor. Another important route is the activation of the Ras / Raf / MEK / ERK cascade. More than 80 substrates are also activated by ERK in the cytoplasm and nucleus. Activated ERK can directly phosphorylate various transcription factors such as Ets, Elk and Myc. Several studies have reported cross-negative and positive control between MEK / ERK and AKT / mTOR, thus affecting MEK / ERK and AKT / mTOR (MC Mendoza et al. , Mol Cell . 18; 41 (6): 661-671, 2011). It has also been shown that AKT and ERK1 / 2 phosphorylate one of the two subunits of TSC1 / TSC2, interfering with this interaction and activating mTOR. Activation of mTOR induces the activity of S6K1 and phosphorylates 4E-BP1, phosphorylated 4E-BP1 separates from eIF4E and becomes active, enhancing cap-dependent translation of mRNA (Liwei Rong et al . , RNA , 14 (7): 1318-27, 2008).

In the present invention, the ability of regenerating the cells of chamomile hydrosol and campo is confirmed by confirming the increase of cell survival and migration according to the treatment of the camomile hydrosol extracted with the vacuum distillation and the main component thereof, campo.

Korean Patent No. 1175803 discloses a composition for improving skin cell regeneration and wrinkles, and Korean Patent No. 0858629 discloses a composition for enhancing skin vitality containing an extract of Wanbai, Haesong, and Baesong extract as an active ingredient. Cosmetic composition " However, as described in the present invention, a composition for skin regeneration containing a campo or chamomile reduced-pressure distillation extract as an active ingredient has not been disclosed.

SUMMARY OF THE INVENTION The present invention has been made in view of the above-mentioned needs, and it is an object of the present invention to provide a method for producing chomomile hydrosol, which comprises extracting chamomile hydromolecules by vacuum distillation to induce cell proliferation and migration, The present inventors have completed the present invention by developing a composition for regenerating skin having excellent skin regeneration ability and in vivo safety containing an extract of chamomile decompression distillate containing as an active ingredient camphor or camphor as a main component Respectively.

In order to solve the above-described problems, the present invention provides a method for producing an active ingredient, which comprises the step of mixing camphor (1R) -1,7,7-trimethylbicyclo [2.2.1] heptan-2-one or camomile hydrosol comprising the same, And a composition for skin regeneration.

The present invention also provides a cosmetic composition for skin regeneration, which contains Campo or a camomile hydrosol containing the same as an active ingredient.

The present invention also provides a pharmaceutical composition for skin regeneration comprising as an active ingredient camphor, a pharmaceutically acceptable salt thereof or a camomile hydrosol comprising the camphor.

The present invention also provides a food composition for regeneration of skin containing Campo or a camomile hydrosol containing the same as an active ingredient.

The composition of the present invention contains a chamomile reduced-pressure distillation extract containing a campo or a camphor as a main component, as an active ingredient, so that it is excellent in skin regeneration ability for promoting the proliferation and migration of skin cells and contains only components derived from a natural herb plant It has no side effects and can be used in various forms such as medicines, quasi-drugs, cosmetics and food ingredients.

Figure 1 shows cell viability and mobility after treatment with vacuum distillation extract (CHV) or distillation extract (CHD) of chamomile. (A) Cell viability of fibroblasts was measured by MTT assay for 24 hours. (B) Cell viability of horny cells (HaCat) was measured. (C) (wound-healing assay), the percentage of cell mobility was measured at 8 sites of each concentration. (D) Distillation extract (CHD; upper) and vacuum distillation extract (CHV; lower).
Figure 2 shows GC-MS results of CHV and CHD. The hydrosol sample was identified by GC-MS apparatus in 10% of CHV (A) and CHD (B).
Figure 3 shows that camo induces migration of HaCat cells and proliferation of fibroblasts. (A) fibroblasts, (B) HaCat cells, (C) HaCat cells.
Figure 4 shows that proliferation of fibroblasts is activated by PI3K / AKT and MAP kinase dependent on ROS production. (A) ROS production of fibroblasts measured by flow cytometry analysis after campo treatment, (B) cell cycle measured by flow cytometry, (C) after treatment of fibroblasts with campo , Phosphorylated AKT, -ERK, -PI3K, -p21, -cyclin D, -cdc-2 and β-actin specific antibodies.
Figure 5 shows that migration of HaCat cells is activated through Vimentin, Slug and Snail, which are regulated by ROS. (A) ROS production of HaCat cells measured by flow cytometry analysis after campo treatment, (B) treatment of Campo with HaCat cells, followed by specific treatment of visenter, slug, snail and β-actin Western blot using antibody.

In order to accomplish the object of the present invention, the present invention provides a process for the preparation of camphor (1R) -1,7,7-trimethylbicyclo [2.2.1] heptan-2-one or camomile hydrosol Which comprises as an active ingredient, a composition for skin regeneration.

In one embodiment of the present invention, a skin regeneration composition containing campo or a camomile hydrosol containing the same as an active ingredient may induce cell proliferation and migration, and the cell is preferably a fibroblast, Or keratinocyte (HaCat), but are not limited thereto.

In one embodiment of the present invention, the cannula may be contained in an amount of 0.0005 to 0.004% by weight based on the total weight of the composition, but is not limited thereto. If the content of the camphor is less than 0.0005% by weight, the effect of promoting skin regeneration by the above-mentioned ingredients can not be obtained. If the content of the camphor is more than 0.004% by weight, the increase of the effect is not so significant.

In one embodiment of the present invention, the chamomile hydrosol may be prepared by distilling the chamomile, preferably by vacuum distillation, more preferably by vacuum extraction at a pressure of 95-100 kPa, But it is not limited thereto. The vacuum distillation method is a general distillation method known in the art and is not particularly limited as far as the distillation method is such that the content of the camphor in the chamomile hydrosol is at least 60% by weight based on the total weight of the hydrosol.

In one embodiment of the present invention, the chamomile hydrosol may be contained in an amount of 1.0 to 20% by weight based on the total weight of the composition, but is not limited thereto. If the content of the camomile hydrosol is less than 1.0% by weight, the skin regeneration promoting effect by the above-mentioned ingredients can not be obtained. If the content exceeds 20% by weight, the increase of the content of the camomile hydrosol is not so large, which is rather inefficient. The chamomile hydrosol preferably includes, but is not limited to, a camphor content of 60 wt% or more, preferably 60 to 90 wt%, based on the total weight of the hydrosol.

The present invention also provides a cosmetic composition for skin regeneration, which contains Campo or a camomile hydrosol containing the same as an active ingredient.

The cosmetic composition for skin regeneration according to the present invention can be used as a skin regenerating cosmetic composition for skin regeneration, such as softening longevity, convergent lotion, nutritional lotion, nutritional cream, massage cream, essence, eye cream, eye essence, cleansing cream, cleansing foam, cleansing water, pack, powder, , Body oils, and body essences, but are not limited thereto.

The cosmetic composition may further contain, in addition to the campo-hydrate of the present invention or the camomile hydrosol containing the same, a lipid, an organic solvent, a solubilizer, a thickening agent and a gelling agent, a softener, an antioxidant, a suspending agent, a stabilizer, , A perfume, a surfactant, water, an ionic or nonionic emulsifier, a filler, a sequestering and chelating agent, a preservative, a vitamin, a barrier, a wetting agent, a essential oil, a dye, a pigment, a hydrophilic or lipophilic active agent, Or any other ingredient conventionally used in cosmetics. ≪ RTI ID = 0.0 > [0040] < / RTI >

The present invention also provides a pharmaceutical composition for skin regeneration comprising as an active ingredient camphor, a pharmaceutically acceptable salt thereof or a camomile hydrosol comprising the camphor.

The pharmaceutical composition according to the present invention can be used as an active ingredient in a medicament in the form of a pharmaceutically acceptable salt thereof. The salt is not particularly limited as long as it is pharmaceutically acceptable so long as it is pharmaceutically acceptable and includes, for example, hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, hydrofluoric acid, hydrobromic acid, formic acid acetic acid, tartaric acid, lactic acid, citric acid, fumaric acid, , Benzenesulfonic acid, toluenesulfonic acid, and naphthalenesulfonic acid. In addition to acid addition salts, additional base salts such as sodium hydroxide, potassium hydroxide, triethylamine, tertiary-butylamine may also be used.

In the formulation of the pharmaceutical composition according to the present invention, the content of the active ingredient may vary widely depending on the formulations, and is generally according to a conventional method.

The pharmaceutical composition according to the present invention may be formulated by adding various adjuvants such as carriers and other additives such as stabilizers, emollients, emulsifiers and the like, which are used in medicines, if necessary, so long as they do not adversely affect the active ingredients.

In addition, the composition according to the present invention can be administered parenterally or orally. As the route of parenteral administration, transdermal administration is preferable, and local application is most preferable. For example, it may be manufactured in the form of a bandage, but is not limited thereto. The formulations may be any formulations suitable for pharmaceutical preparations including ointments, creams, injections, powders, granules, tablets, and the like.

The preferred dosage of the pharmaceutical composition according to the present invention may be 0.001 to 1000 mg / Kg · day, but is not limited thereto. In addition, the composition according to the present invention may be administered alone, or may be administered together with other medicines in order to aid in the same or another medicament. In addition, in the composition of each formulation, components other than the composition, which is the above-mentioned essential ingredient, can be mixed and selected by a person skilled in the art according to the formulation or purpose of use of the other external preparation. In this case, Can happen.

The present invention also provides a food composition for regeneration of skin containing Campo or a camomile hydrosol containing the same as an active ingredient.

When the composition of the present invention is used as a food additive, the composition may be added as it is, or may be used together with other food or food ingredients, and suitably used according to a conventional method. The amount of the active ingredient to be mixed can be suitably determined according to its intended use (prevention, health or therapeutic treatment). Generally, the composition of the present invention is added in an amount of not more than 15 parts by weight, preferably not more than 10 parts by weight, based on the raw material, when the food or beverage is produced. However, in the case of long-term consumption intended for health and hygiene purposes or for health control purposes, the amount may be less than the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount exceeding the above range .

There is no particular limitation on the kind of the food. Examples of foods to which the above substances can be added include dairy products including meats, sausages, breads, chocolates, candies, snacks, confectionery, pizza, ramen noodles, gums, ice cream, soups, drinks, tea, Alcoholic beverages, and vitamin complexes, all of which include healthy foods in a conventional sense.

The health beverage composition of the present invention may contain various flavors or natural carbohydrates as an additional ingredient such as ordinary beverages. Such natural carbohydrates are monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, and polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol and erythritol. Examples of sweeteners include natural sweeteners such as tau martin and stevia extract, synthetic sweeteners such as saccharin and aspartame, and the like. The ratio of the natural carbohydrate is generally about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g per 100 ml of the composition of the present invention.

In addition to the above, the composition of the present invention may further comprise various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and salts thereof, alginic acid and its salts, organic acids, protective colloid thickening agents, pH adjusting agents, stabilizers, preservatives, glycerin, A carbonating agent used in beverages, and the like. In addition, the composition of the present invention may contain flesh for the production of natural fruit juices, fruit juice drinks and vegetable drinks. These components may be used independently or in combination. The proportion of such additives is not critical, but is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.

Hereinafter, the present invention will be described in detail by way of examples. However, the following examples are for illustrative purposes only and are not intended to limit the scope of the present invention.

Materials and methods

1. Reagents

RPMI 1640, DMEM medium, bovine serum albumin (BSA), trypsin / EDTA, fetal bovine serum (FBS), antibiotics and Hoechst 33342 were purchased from Invitrogen. MTT (3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazoliumbromide) was purchased from Amresco and DCFH-DA (2'-7'-Dichlorodihydrofluorescein diacetate) was purchased from Sigma. Anti-PI3K, -AKT, -4E-BP1, -ERK, -Vimentin, -Slug, -Snail, -β-actin antibodies were purchased from Cell signaling and PVDF (Polyvinylidene fluoride) membranes were purchased from Bio-Rad.

2. Samples and Hydrosol  extraction

Chamomile (chamomile) samples grown produce in ㈜ Urban Farmer's farm is located in Seogwipo, Jeju Island pyoseonmyeon German chamomile (Matricaria recutita L.) and a 30 cm flowering stalk were collected and used in the summer of 2012. The raw samples were extracted by distillation or vacuum extraction at 95 kPa-100 kPa. The evaporated and condensed hydrozols were recovered and stored in a refrigerator.

3. Cell culture

Human epidermal specimens were obtained from 8 donors between 6 and 12 years old who were circumcised at Jeju University Hospital. The specimens were washed with sterile PBS and the subcutaneous tissue was carefully removed. The skin was then cut into small pieces of 1-2 mm ³ . After treatment with 0.1% dispose at 4 ° C overnight, the epithelial layer was removed, and the remaining skin was treated with 0.1% collagenase I at 37 ° C for 4 hours. The enzyme-treated cells were passed through a 70-mm cell filter (BD Biosciences, Mississauga, ON, Canada), centrifuged and resuspended in DMEM medium containing 10% FBS, 100 U / ml penicillin, 100 μg / ml streptomycin Lt; / RTI > The cells were inoculated at 1 × 10 ³ cells / cm ² in a tissue culture dish and cultured at 37 ° C and 5% CO ₂ . After 24 hours, the culture dish was washed with PBS and unattached cells were removed. The remaining adherent cells were proliferated until they became dense within 7 days. The keratinocyte (HaCat) and dermal fibroblast were cultured in RPMI 1640 medium containing 10% FBS and 1% antibiotics, and DMEM medium at 37 ° C and 5% CO _{2, respectively} .

4. Measurement of cell viability

The keratinocytes and fibroblasts were subcultured to a microtiter plate at 2 × 10 ⁴ cells / ml. After 48 hours, various concentrations of chamomile hydrosol were treated and incubated again for 48 hours. After the incubation, 20 μl of MTT solution (5 mg / ml) was added and incubated for 4 hours. The supernatant was removed and 150 μl of DMSO was added. Cell viability was determined by measuring the absorbance at 570 nm using a Sunrise microplate reader (Tecan, Salzburg, Austria).

5. Flow cytometry

To measure reactive oxygen species (ROS), the cells were divided into 2 × 10 ⁴ cells / ml in a 60 mm culture dish and treated with 10% chamomile hydrosol for 6, 12, and 24 hours. The cells were stained with DCFH-DA (2'-7'-Dichlorodihydrofluorescein diacetate) for 15 minutes, and the cells were collected in a 15 ml tube and washed with PBS. The cells were resuspended in 500 [mu] l of PBS and the level of ROS was measured. For cell cycle analysis, the treated cells were collected and fixed in 70% ethanol at 4 ° C overnight. After 24 hours, the cells were rehydrated with 2 mM EDTA-PBS containing RNase A (25 ng / ml) and stained with PI (40 μg / ml). All analyzes were performed using a FACS Caliber flow cytometer (BD Biosciences) and each sample was analyzed by measuring 10,000 cells.

6. Gas Chromatography-Mass Spectrometry ( GC - MS )

Gas chromatographic analysis was performed using Shimadzu GC-MS (Model QP-2010, Shimadzu Co., Kyoto, Japan) (electron impact, ionization voltage 70 eV). A GC column (30 m long, 0.25 mm internal diameter and 0.25 μm film thickness) was used with Rtx-5MS.

7. Cell migration analysis or wound healing analysis ( In vitro wound healing assay )

The cells were dispensed into 6-well plates and after 24 hours the cells were scraped off and the medium removed when the cell density reached about 80%. After addition of fresh medium, the cells were treated with 0-20% concentration of chamomile hydrosol or 0-40 [mu] g / ml of campo and allowed to react for 32 hours. After 32 hours, the cells were observed under a microscope, and the degree of cell migration was compared with that of the control group. The average value was obtained by measuring 8 samples of each sample.

8. Western Blot  analysis

Cells were harvested at 3, 6, 12 and 24 hours after treatment with 0-20% concentration of camomile hydrosol or 0-40 μg / ml of campo and incubated in cell lysis buffer (100 mM Tris-HCl pH 8, 250 mM NaCl , 0.5% Nonidet P-40 and 1x protease inhibitor cocktail) and left on ice for 30 minutes. Cell lysates were centrifuged at 13,000 rpm for 30 min at 4 ° C to separate supernatant, and then the concentration was measured with a BCATM Protein assay (Pierce, Rockford, IL). Cell lysates were separated by 10-15% SDS-PAGE and transferred to PVDF membranes using glycine transfer buffer (192 mM glycine, 25 mM Tris-HCl (pH 8.8) and 20% (v / v) methanol). Membranes were blocked with 5% skim milk overnight at 4 ° C and then reacted with primary antibody for 1 hour at room temperature or overnight at 4 ° C, and then treated with secondary antibody for 1 hour. The primary antibody was diluted at a ratio of 1: 1000, and the anti-β-actin antibody and the secondary antibody were diluted at a ratio of 1: 10,000. Protein bands were analyzed using WEST-ZOL® plus Western Blot Detection System (iNtRON, Gyeonggi-do, Korea).

9. Statistics

Statistical analysis was performed by one-way analysis (ANOVA) and Student's t-test using SPSS v.12.0 software, and flow cytometry analysis was performed in three replicates.

Example  One. Hydrosol  Main ingredient Campo  Survival of fibroblasts and migration of keratinocytes

The skin includes various types of cells such as fibroblasts, keratinocytes and melanocytes. To evaluate the effect of chamomile hydrosol on skin, keratinocytes (HaCat) and fibroblasts were used. As shown in Figure 1A, 10% CHV (chamomile hydroisol extracted with reduced pressure distillation) showed a proliferation effect of 10% or more in fibroblast. However, no proliferation effect was observed in keratinocytes (FIG. 1B). In addition, CHV showed high efficacy in migration of keratinocytes. In particular, treatment with 10% CHV showed a double increase compared to the control. Compared with the two methods of vacuum distillation-extracted chamomile hydroisomer (CHV) and distilled-extracted chamomile hydrosol (CHD), CHV induced cell proliferation and migration, but CHD did not induce cell proliferation and migration , B, C). This means that the content of important components affecting the activity of the hydrosol is different depending on the extraction process. For this reason, the components of CHV and CHD were analyzed using gas chromatography-mass spectrometry. Gas Chromatography-Mass spectrometry results showed that the CHV contained 68.11% of the campo, while the CHD contained only 0.51% (Figures 2A, B and 1A, B). This suggests that Camphor is a major component of cell migration and proliferation.

Since the camo is contained in a relatively large amount in CHV, the activity of CHV on fibroblasts and keratinocytes was measured. The results of MTT analysis showed that cell viability of fibroblasts was increased when camogue was treated at 5 ㎍ / ㎖ to 40 ㎍ / ㎖, but the survival rate of keratinocytes was not increased (Fig. 3A, B). However, the results of the wound healing analysis showed that migration of keratinocytes was significantly increased when the camber was treated at 20 / / ㎖ to 40 / / ㎖ (Fig. 3C).

These results show that CHV containing more than 60% of the cancer cells has an activity of inducing cell proliferation and migration, but CHD having almost no cancer cell shows little cell proliferation and migration activity. Therefore, it can be concluded that CHV induces cell proliferation and migration because it contains campo as a main component.

Example  2. Campo  By treatment Active oxygen species  ( ROS )

Generation of reactive oxygen species (ROS) is known to be a stress that causes DNA damage or various cellular diseases, and is associated with various cell deaths such as cell proliferation, self-extinguishing action, and necrosis. However, reactive oxygen species have a good side that plays an important role in cell signaling and regulation. Since the increase of reactive oxygen species is an index of cell proliferation, division and migration, the production level of reactive oxygen species by kerosene treatment was measured in keratinocytes and fibroblasts. As a result, in the fibroblasts, the production of reactive oxygen species was significantly increased after 24 hours from the campo treatment (FIG. 4A). In the keratinocytes, the production of reactive oxygen species was significantly increased from 12 hours to 24 hours after treatment (Fig. 5A). The production of reactive oxygen species increased about 13% and 16% in fibroblasts and keratinocytes, respectively. This level of production is consistent with other reports that endogenous reactive oxygen species can promote cell migration and proliferation at non-toxic levels (Blanchetot and Boonstra, Crit Rev Eukaryot Gene Expr , 18 (1): 35-45, 2008). These results suggest that the signal of cell survival is related to the production of reactive oxygen species and that cellular signals are regulated by reactive oxygen species in these cells.

Example  3. Camper  Induced Reactive oxygen species  By production MAP Kinase , PI3K / AKT  Path control

Signal transduction mediated by reactive oxygen species usually leads to life-sustaining pathways. In order to understand the molecular mechanism by the camphor treatment, it was confirmed that the camo induces the lower signal of reactive oxygen species production. Campoites were treated with fibroblasts over time and Western blotted. As a result of Western blot, it was shown that campo increased the expression of active forms of phosphorylated PI3K, AKT and ERK (Fig. 4C) and decreased inactivity. Analysis of 4E-BP1, a sub-protein of PI3K / AKT and known to inhibit mRNA translation, was also performed. The increase in phosphorylated 4E-BP1 interferes with complex formation with eIF4E and can be regulated by active eIF4E (Fig. 4C). Increased phosphorylated ERK can be involved in this process. Phosphorylated ERK plays a role as an active transcription factor and interacts with mTOR to increase its function.

In addition, Western blot results showed that the cell cycle marker proteins p21, cdc-2 and cyclin D1 increased (FIG. 4C), which is known to be upregulated when the cell cycle is activated for proliferation. This result is consistent with the cell cycle measurement using flow cytometry (Fig. 4B) in which the G1 and S groups associated with cdc-2 and cyclin D1 are increased by the camptothe treatment. These results indicate that camphor induces cell proliferation through PI3K / AKT and MAP kinase dependent on production of reactive oxygen species in fibroblasts.

Example  4. Camper  Induced Vimentin  ( Vimentin ), Snail  ( Snail ) And Slug  Increased mobility of keratinocytes through activation of slug

Because cell migration and proliferation are associated with the production of reactive oxygen species (FIGS. 4A, 5A), major modulators that induce keratinocyte mobility in reactive oxygen species-dependent signals were investigated. Vimentin is a cytoskeletal factor that plays an important role in cell strength and preservation. Can be used as an indicator of metastasis since visenterin is upregulated during cell migration or epithelial-mesenchymal transition. At the same time, the transcription factor, slug and snail, is increased with the visentin. As a result of western blotting, these indicators were increased by campo processing (Fig. 5B). This result is consistent with reports that active oxygen species induce expression by increasing mRNA stability of snail (Dong et al ., Biochem Biophys Res Commun . , 27: 356 (1): 318-321, 2007). These results show that the camo induces cell migration through vimentin, slug and snail controlled by reactive oxygen species in keratinocytes.

Claims (9)

Camomile hydrosol containing 60 to 90% by weight of camphor (1R) -1,7,7-Trimethylbicyclo [2.2.1] heptan-2-one), prepared by vacuum distillation of chamomile, A composition for regenerating skin containing as an active ingredient. The method according to claim 1, wherein the camomile hydrosol comprising the camphor (1R) -1,7,7-trimethylbicyclo [2.2.1] heptan-2-one) Wherein the composition is derived from the composition. delete delete delete The composition for skin regeneration according to claim 1, wherein the chamomile hydrosol is contained in an amount of 1.0 to 20% by weight based on the total weight of the composition. A cosmetic composition for skin regeneration comprising, as an active ingredient, a camomile hydrosol comprising 60 to 90% by weight of camphor produced by vacuum distillation of chamomile. A cosmetic composition for regenerating skin comprising chamomile hydrosol comprising 60 to 90% by weight of camphor produced by vacuum distillation of chamomile as an active ingredient. 60% to 90% by weight of water, prepared by vacuum distillation of chamomile Claims [1] A food composition for regeneration of skin comprising chamomile hydrosol containing a campo as an active ingredient.

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